Bottom Line:
Inflammation is a growing area of research in neurodegeneration.However, bone marrow CD11b(+) cells did not show the same hyper-responsive phenotype as spleen and blood cells.Taken together, these results show significant promise for these mouse models to be used to study targeting innate immune pathways identified in human cells, thereby helping to understand the role the peripheral immune system plays in HD progression.

f0035: Phagocytosis levels in macrophages isolated from HD patients and R6/2 mice. (A) Monocyte-derived macrophages were incubated with green fluorescent latex beads for 1 h prior to analysis by flow cytometry of the percentage of cells taking up the beads. Macrophages were gated in the FCS and SSC channels according to their size, and the FL-1 channel was used to determine the percentage of cells that had phagocytosed particular numbers of beads. (B) HD patient macrophages stimulated with LPS for 24 h showed a significant increase in the percentage of phagocytosing cells. n = 5 control and 5 HD patients. (C) Levels of phagocytosis in LPS stimulated bone marrow-derived macrophages were similar in 12-week old wild-type and R6/2 mice, whilst (D) peritoneal macrophages from 12-week old R6/2 mice stimulated with LPS for 24 h showed a significant increase in phagocytosis levels. n = 5 wild-type and n = 4 R6/2 mice. Unpaired two-tailed Student's t-test.

Mentions:
Phagocytosis is a key function of macrophages and a significant elevation in phagocytosis was observed in LPS-stimulated blood-derived HD patient macrophages compared with control cells in a flow cytometry based assay (Fig. 7B). To see whether R6/2 mice demonstrate a similar phenotype, bone marrow-derived and peritoneal macrophages were incubated with green fluorescent latex beads and the percentage of cells phagocytosing beads were analysed by flow cytometry. LPS-stimulated bone marrow-derived macrophages from 12-week old R6/2 mice did not demonstrate a difference in phagocytosis levels compared with wild-type mice (Fig. 7C). Peritoneal macrophages isolated from R6/2 mice, however, showed a small elevation in the percentage of phagocytosing cells upon LPS stimulation, similar to the results in HD patients (Fig. 7D).

f0035: Phagocytosis levels in macrophages isolated from HD patients and R6/2 mice. (A) Monocyte-derived macrophages were incubated with green fluorescent latex beads for 1 h prior to analysis by flow cytometry of the percentage of cells taking up the beads. Macrophages were gated in the FCS and SSC channels according to their size, and the FL-1 channel was used to determine the percentage of cells that had phagocytosed particular numbers of beads. (B) HD patient macrophages stimulated with LPS for 24 h showed a significant increase in the percentage of phagocytosing cells. n = 5 control and 5 HD patients. (C) Levels of phagocytosis in LPS stimulated bone marrow-derived macrophages were similar in 12-week old wild-type and R6/2 mice, whilst (D) peritoneal macrophages from 12-week old R6/2 mice stimulated with LPS for 24 h showed a significant increase in phagocytosis levels. n = 5 wild-type and n = 4 R6/2 mice. Unpaired two-tailed Student's t-test.

Mentions:
Phagocytosis is a key function of macrophages and a significant elevation in phagocytosis was observed in LPS-stimulated blood-derived HD patient macrophages compared with control cells in a flow cytometry based assay (Fig. 7B). To see whether R6/2 mice demonstrate a similar phenotype, bone marrow-derived and peritoneal macrophages were incubated with green fluorescent latex beads and the percentage of cells phagocytosing beads were analysed by flow cytometry. LPS-stimulated bone marrow-derived macrophages from 12-week old R6/2 mice did not demonstrate a difference in phagocytosis levels compared with wild-type mice (Fig. 7C). Peritoneal macrophages isolated from R6/2 mice, however, showed a small elevation in the percentage of phagocytosing cells upon LPS stimulation, similar to the results in HD patients (Fig. 7D).

Bottom Line:
Inflammation is a growing area of research in neurodegeneration.However, bone marrow CD11b(+) cells did not show the same hyper-responsive phenotype as spleen and blood cells.Taken together, these results show significant promise for these mouse models to be used to study targeting innate immune pathways identified in human cells, thereby helping to understand the role the peripheral immune system plays in HD progression.